Figure 1.

Lack of CREB or C/EBP β expression alters inflammatory responses in macrophages. RAW 264.7 cells were transduced with lentivirus containing Cas9 and gRNA directed against CREB or C/EBP β. Following transduction, clones were selected that do not express CREB (RAW^ΔCREB) or C/EBP β (RAW^{Δtotal C/EBPβ}). Panel (A) Demonstration of loss of CREB expression as shown by immunoblotting for CREB and GAPDH. Panel (B) Immunoblot analysis of C/EBP β and GAPDH expression in RAW^ΔCREB cells and control cells after exposure to ET (10 nM of EF and 10 nM of PA) for 6 h. Panel (C) RAW^{Δtotal C/EBPβ} cells lack C/EBP β expression as shown by immunoblot analysis of CREB, C/EBP β, and GAPDH. Panel (D–G) RAW^ΔCREB cells, RAW^{Δtotal C/EBPβ} cells, and control RAW 264.7 cells were exposed to 1 μg/ml LPS for 6 h. RT-qPCR was used to quantify transcript levels of Il6, TNFa, Inos, and Il10. RT-qPCR data are presented as mean (n = 3) ± S.D. Asterisks indicate significant different than control cells. *p < 0.05.